METHOD FOR PURIFYING HIV REVERSE TRANSCRIPTASE

摘要

Relating to the production and purification of recombinant proteins, a process for purifying recombinant reverse transcriptase from human immunodeficiency virus (HIV-RT) is provided. E. coli lysate containing HIV-RT is contacted with polyethyleneimine to precipitate cell debris and nucleic acids; the resulting supernatant is contacted with anion exchange resin at low salt, pH 6-9; the eluate is contacted with a cation exchange resin at pH 6-8; the second eluate is contacted with a second anion exchange resin to remove an E. coli protease; and the third eluate is contacted with a third anion exchange resin at a pH above 8.8 to yield highly purified, protease-free HIV-RT.